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Adjuvant for enhancing fish vaccine immunization effect and application thereof

An immunization and adjuvant technology, applied in the field of adjuvants that can enhance the effect of fish vaccine immunization, can solve problems such as no research and application reports, achieve harmlessness to the environment and operators, and improve specific and non-specific immunity. The effect of protection rate and safe usage

Active Publication Date: 2012-05-02
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, astragalus extract as an adjuvant to enhance fish vaccine immunization has no research and application reports at home and abroad.

Method used

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  • Adjuvant for enhancing fish vaccine immunization effect and application thereof
  • Adjuvant for enhancing fish vaccine immunization effect and application thereof
  • Adjuvant for enhancing fish vaccine immunization effect and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Determination of safe dosage of astragaloside for zebrafish injection

[0025] 1. Materials and methods:

[0026] (1) Experimental fish and breeding management: Healthy zebrafish (0.32±0.08g) were selected and placed in separate breeding tanks (10L / tank, Shanghai Haisheng Aquarium Equipment Factory) with a water temperature of 28±1°C. The daily lighting is 12h. During the test period, the fish were fed with compound feed at 3% of their body weight, twice a day, and kept for 2 weeks before the test. The zebrafish were soaked and anesthetized with MS222 (diluted 1:10000) before injection.

[0027] (2) Determination of the safe dose of astragaloside for zebrafish injection: 20 zebrafish were intraperitoneally injected with astragaloside at a concentration of 1, 2, 5, 8, 10, 12, 15 and 20 mg / mL, respectively, and the injection dose was 10 μL / Tail, observed for 14 days.

[0028] 2. Test results

[0029] The death of zebrafish in each experimental group withi...

Embodiment 2

[0032] Example 2: Astragalus saponin was added to formalin-inactivated E. tarda vaccine, and zebrafish (Danio rerio) were immunized by injection.

[0033] 1. Materials and methods:

[0034] (1) Test fish and breeding management: same as embodiment 1.

[0035] (2) Vaccine preparation: inoculate E.tarda in tryptone soy broth (TSB), culture with shaking at 28°C overnight, then transfer to fresh TSB at a ratio of 1:10, culture with shaking at 28°C for 5 hours, collect the bacterial liquid, 4000g , 4°C, 10 min, wash with sterile PBS (pH7.2) for 3 times, resuspend in PBS, and spread on Tryptone Soy Agar (TSA) after dilution. 9 CFU / mL bacterial suspension, placed in 1% formalin (36.5-38%, w / v), overnight at 4°C to make an inactivated vaccine, washed 3 times with PBS (the method is the same as above), and stored at 4°C . Another 0.2 mL of the inactivated bacterial solution was coated with TSA, and cultured at 28°C for 48 hours to confirm that there was no colony growth. In the vac...

Embodiment 3

[0047] Example 3: Determination of safe dosage of astragaloside for turbot injection

[0048] 1. Materials and methods:

[0049] (1) Test fish and breeding management: select healthy turbot (11.4 ± 1.8g), temporarily raise it in a cement pond for 2 weeks before the test, and place it in a round glass fiber reinforced plastic bucket (water 1 meter) 3 ), the water temperature is 18±2°C, the salinity is 28‰±2‰, continuous aeration for 24 hours, and running water culture. During the test period, the compound feed was fed according to 3% of the fish body weight, 3 times a day, and the water was changed and cleaned 2 times a day. The turbot was soaked and anesthetized with MS222 (1:9000) before injection.

[0050] (2) Determination of the safe dose of astragaloside for turbot injection: 20 turbots were intraperitoneally injected with astragaloside with a concentration of 2, 5, 8, 10, 15 and 20 mg / mL respectively, and the injection dose was 100 μL / tail. Observed for 14 days.

[0...

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Abstract

The invention relates to an adjuvant for enhancing a fish vaccine immunization effect and an application thereof. The adjuvant for enhancing the fish vaccine immunization effect is characterized in that an immune potentiator is extracted from Astragalus mongholicus as a leguminous plant or effective components of the Astragalus mongholicus, comprising Astragaloside and Astragalus polysacharin, and natural products or manually modified products or manually synthetic products comprising the Astragaloside and the Astragalus polysacharin can be adopted to serve as the effective components. The adjuvant is capable of increasing the specific immune protection rate after vaccine component immunization is finished. When the adjuvant is applied, a vaccine can comprise the following components: any one or more than one of expression products of inactivated pathogens, bacterial ghost components, hypotoxic pathogens, attenuated pathogens, protective antigens, antigen subunits, antigen determinants or antigen gene expression vectors of bacteria, viruses and parasites. When in use, the adjuvant can be mixed with the components of the vaccine for application, also cannot be mixed with the components of the vaccine for application and also can be applied with the vaccine at different time.

Description

technical field [0001] The invention relates to fish vaccine immunization prevention and control technology, in particular to an adjuvant capable of enhancing the effect of fish vaccine immunization and its application. Background technique [0002] The pathogens of fish infectious diseases mainly include bacteria, viruses or parasites, which can cause acute, subacute or chronic infection of fish, causing aquaculture production to be attacked by diseases, causing serious losses and seriously endangering the sustainable development of the aquaculture industry . The use of antibiotics or chemical drugs for prevention or treatment often brings disadvantages such as microbial resistance, drug residues in products, and environmental pollution. Fish vaccine immunization against infectious diseases can safely and efficiently control the occurrence and prevalence of diseases , is an internationally accepted disease control technology, and is of great significance to the healthy and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61P37/04
Inventor 黄倢谢国驷王秀华隋虎辰边慧慧张庆利宋晓玲刘莉
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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